Therapeutic compositions and methods

ABSTRACT

The present invention relates in general to therapeutic compositions and methods of use. In preferred embodiments, the invention relates to the field of eye health. In some embodiments, the invention relates to the prevention and treatment of macular degeneration by administering compounds disclosed herein. In some embodiments, the invention relates to compositions and methods of improving vision.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.13/304,088, filed Nov. 23, 2011, which is a divisional of U.S. patentapplication Ser. No. 11/416,773, filed May 3, 2006, now U.S. Pat. No.8,088,773, which claims priority to U.S. Provisional Application No.60/680,998, filed May 12, 2005 and U.S. Provisional Application No.60/776,426, filed of Feb. 24, 2006; this application is also acontinuation of U.S. patent application Ser. No. 13/304,116, filed Nov.23, 2011, which is a divisional of U.S. patent application Ser. No.11/416,773, filed May 3, 2006, now U.S. Pat. No. 8,088,773, which claimspriority to U.S. Provisional Application No. 60/680,998, filed May 12,2005 and U.S. Provisional Application No. 60/776,426, filed of Feb. 24,2006, all of which are incorporated by reference herein in theirentirety.

TECHNICAL FIELD

The present invention relates in general to therapeutic compositions andmethods of use. In preferred embodiments, the invention relates to thefield of eye health. In some embodiments, the invention relates to theprevention and treatment of macular degeneration by administeringcompounds disclosed herein. In some embodiments, the invention relatesto compositions and methods of improving vision.

BACKGROUND

Aging is a chronic process causing degeneration of cells, tissues, andorgans, including choroidal blood vessels, retinal pigment epitheliumcells (RPEC) and Bruch's membrane of the eye. Arteriosclerotic agingchanges choroids blood vessels, particularly the macularchorio-capillaris with a decrease in total capillary membrane bloodflow. As a result, retinal pigment epithelium starts to accumulatelipofuscin, alters cell shape, density, pigmentation, lysosomal activityand extracellular matrix formation. Gradually, Bruch's membrane showsthickening and decreased permeability, resulting with breakdown thatallows choroidal neovascularization (CNV) to appear ultimately resultingin age-related macular degeneration and blindness. Thus, there is a needto identify agents that prevent choroidal neovascularization.

BRIEF SUMMARY

The present invention relates in general to therapeutic compositions andmethods of use. In preferred embodiments, the invention relates to thefield of eye health. In some embodiments, the invention relates to theprevention and treatment of macular degeneration by administeringcompounds disclosed herein. In some embodiments, the invention relatesto compositions and methods of improving vision.

Ischemia of choroidal blood flow is one of the major causes ofage-related macular degeneration (AMD). Therefore, agents have beendiscovered to prevent AMD formation via increasing of choroidal bloodflow as measured with colored micro-sphere technique, retinal functionrecovery after ischemic insult, and inhibition of choroidalneovascularization in a laser treated rat model. These agents include:hypotensive agents, such as hydralazine, guanabenz, and D-timolol;flavonoids, such as apigenin, naringenin, quercetin, and flavon; andN-nitropyrazoles and C-nitro-pyrazoles, such as DN6, DN7, DN13, andDC-5. As reduction of choroidal neovascularization (CNV) serves as themajor mechanism to prevent/treat AMD, agents, as demonstrated herein,prevent AMD by preventing or reducing CNV.

In some embodiments, the invention relates to methods of identifying acompound capable of treating an eye disease, preferably maculardegeneration, using methods disclosed herein. Preferred methods includeproviding ischemic insult, measuring retinal function recovery, andcorrelating inhibition of neovascularization to a compound effective forpreventing or treating eye diseases.

In one embodiment, the effective amount of the agent is between 0.1 and250 mg/kg of the patient's weight, depending on the amount of activeagent required and as taught herein. The agent may be adapted for oral,parenteral, intravenous, topical, intracameral or intraocularadministration. In one embodiment, the agent is provided in dry form,e.g., lyophilized and may be resuspended using, e.g., saline, bufferedsaline and the like. The agent may be combined with a suitable carrier,e.g., an anionic, mucomimetic polymer; a gelling polysaccharide; afinely-divided drug carrier substrate; a mineral oil; a liquidpetrolatum; a white petrolatum; a propylene glycol; a polyoxyethylene; apolyoxypropylene; an emulsifying wax; water and mixtures andcombinations thereof.

The present invention also includes a method for treating, preventing ormanaging age-related macular degeneration by administering to a subjectin need thereof a composition that includes an effective amount of anagent that increases choroidal blood flow. Generally, the compositionmay be administered after the occurrence of an acute ischemic traumaevent, may be used before any symptoms are visible or detectable, duringand/or after ischemic trauma.

The subject may be any mammal, e.g., the subject may be a human. Theeffective amount of the agent for pre-treatment, treatment or even postoperative treatment (when used in conjunction with physical interventionand/or other pharmacological therapy) may be determined by measuring oneor more retinal and/or choroidal functions during the recovery after anischemic insult. Depending on the one or more agents selected fortreatment, these may be provided to maximize the effectiveness of theagent, e.g., the effective amount of the agent may be between about 0.1to about 250 mg/Kg depending on the patient's current and future needsfor treatment.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising compounds disclosed herein or salt thereof and substitutedand unsubstituted derivatives thereof functioning to decrease choroidalneovascularization and 2) administering said compound to said subject.In further embodiments, said eye disease is age-related maculardegeneration. In further embodiments, said subject is a human. Infurther embodiments, said administration is topically to the eye. Infurther embodiments, said salt is a hydrochloride salt. In furtherembodiments, said composition is a liquid solution.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising compounds disclosed herein or salt thereofand substituted and unsubstituted derivatives thereof functioning todecrease choroidal neovascularization and 2) administering said compoundto said subject. In further embodiments, said eye disease is age-relatedmacular degeneration. In further embodiments, said subject is a human.In further embodiments, said administration is topically to the eye. Infurther embodiments, said salt is a hydrochloride salt. In furtherembodiments, said composition is a liquid solution.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising tetramethylpyrazine or salt thereof and 2)administering said compound to said subject. In further embodiments,said macular degeneration is age-related. In further embodiments, saidsubject is a human. In further embodiments, said administration istopically to the eye. In further embodiments, said salt is ahydrochloride salt. In further embodiments, said composition is a liquidsolution.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed or at risk for macular degeneration and ii) a compositioncomprising tetramethylpyrazine or salt thereof and 2) administering saidcompound to said subject. In further embodiments, said maculardegeneration is age-related. In further embodiments, said subject is ahuman. In further embodiments, said administration is topically to theeye. In further embodiments, said salt is a hydrochloride salt. Infurther embodiments, said composition is a liquid solution.

In some embodiments, the invention relates to a method of treating orpreventing an eye disease comprising: 1) providing i) a subject and ii)a composition comprising agent or salt thereof and 2) administering saidcompound to said subject. In further embodiments, said eye disease ismacular degeneration. In further embodiments, said subject is a human.In further embodiments, said administration is topically to the eye. Infurther embodiments, said salt is a hydrochloride salt. In furtherembodiments, said composition is a liquid solution. In furtherembodiments, said agent composition is greater than 50%, 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, 99.5% or 99.9% by weight of anisomeric component. In further embodiments, the subject is diagnosed orat risk for macular degeneration. In further embodiments, said subjectexhibits a symptom of macular degeneration, and in further embodimentssaid administering causes a reduction in said symptom

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising timolol or salt thereof and 2) administering said compound tosaid subject. In further embodiments, said macular degeneration isage-related. In further embodiments, said subject is a human. In furtherembodiments, said administration is topically to the eye. In furtherembodiments, said salt is a hydrochloride salt. In further embodiments,said composition is a liquid solution. In further embodiments, saidtimolol composition is greater than 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, 98%, 99%, 99.5% or 99.9% by weight of a D-timololcomponent.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising timolol or salt thereof and 2)administering said compound to said subject. In further embodiments,said macular degeneration is age-related. In further embodiments, saidsubject is a human. In further embodiments, said administration istopically to the eye. In further embodiments, said salt is ahydrochloride salt. In further embodiments, said composition is a liquidsolution. In further embodiments, said timolol composition is greaterthan 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, 99.5%or 99.9% by weight of a D-timolol component.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted compound of the followingformula:

or salt thereof functioning to decrease choroidal neovascularization and2) administering said compound to said subject.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted compound ofthe following formula:

or salt thereof functioning to decrease choroidal neovascularization and2) administering said compound to said subject.

In additional embodiments, the invention relates to a method of treatingor preventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising hydralazine or salt thereof and 2) administering saidcompound to said subject. In further embodiments, said maculardegeneration is age-related. In further embodiments, said subject is ahuman. In further embodiments, said administration is topically to theeye. In further embodiments, said salt is selected from the groupconsisting of a hydrochloride salt, hydrochlorothiazide salt orisosorbide dinitrate salt. In further embodiments, said composition is aliquid solution.

In additional embodiments, the invention relates to a method of treatingor preventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising hydralazine or salt thereof and 2)administering said compound to said subject. In further embodiments,said macular degeneration is age-related. In further embodiments, saidsubject is a human. In further embodiments, said administration istopically to the eye. In further embodiments, said salt is selected fromthe group consisting of a hydrochloride salt, hydrochlorothiazide saltor isosorbide dinitrate salt. In further embodiments, said compositionis a liquid solution.

In another embodiment, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subject asubject diagnosed with or at risk for macular degeneration and ii) acomposition comprising a substituted or unsubstituted compound of thefollowing formula:

or salt thereof functioning to decrease choroidal neovascularization and2) administering said compound to said subject.

In another embodiment, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subject asubject that exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted compound ofthe following formula:

or salt thereof functioning to decrease choroidal neovascularization and2) administering said compound to said subject.

In another embodiment, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising guanabenz or salt thereof and 2) administering said compoundto said subject. In further embodiments, macular degeneration isage-related. In further embodiments, said subject is a human. In furtherembodiments, said administration is topically to the eye. In furtherembodiments, said salt is selected from the group of a hydrochloridesalt and an acetate salt. In further embodiments, said composition is aliquid solution.

In another embodiment, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising guanabenz or salt thereof and 2)administering said compound to said subject. In further embodiments,macular degeneration is age-related. In further embodiments, saidsubject is a human. In further embodiments, said administration istopically to the eye. In further embodiments, said salt is selected fromthe group of a hydrochloride salt and an acetate salt. In furtherembodiments, said composition is a liquid solution.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted compound of the followingformula:

or salt thereof functioning to decrease choroidal neovascularization and2) administering said compound to said subject.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted compound ofthe following formula:

or salt thereof functioning to decrease choroidal neovascularization and2) administering said compound to said subject.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subject asubject diagnosed with or at risk for macular degeneration and ii) acomposition comprising a compound having the following structure:

or salt thereof wherein, R¹ is hydrogen, alkyl, aryl, or arylalkyl; R²and R³ are the same or different and, at each occurrence, independentlyhydrogen, alkyl, or alkylcarboxyl; R² and R³ together form a fivemembered lactone ring; R⁴ is hydrogen or alkyl; and R⁵ is alkyl; and 2)administering said compound to said subject. In further embodiments,said macular degeneration is age-related. In further embodiments, saidsubject is a human. In further embodiments, said administration istopically to the eye. In further embodiments, said salt is ahydrochloride salt. In further embodiments, said composition is a liquidsolution.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subject asubject that exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a compound having the following structure:

or salt thereof wherein, R¹ is hydrogen, alkyl, aryl, or arylalkyl; R²and R³ are the same or different and, at each occurrence, independentlyhydrogen, alkyl, or alkylcarboxyl; R² and R³ together form a fivemembered lactone ring; R⁴ is hydrogen or alkyl; and R⁵ is alkyl; and 2)administering said compound to said subject. In further embodiments,said macular degeneration is age-related. In further embodiments, saidsubject is a human. In further embodiments, said administration istopically to the eye. In further embodiments, said salt is ahydrochloride salt. In further embodiments, said composition is a liquidsolution.

In further embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted compound of the followingformula:

or salt thereof functioning to decrease choroidal neovascularizationwherein, R¹ is hydrogen, alkyl, substituted alkyl, aryl, substitutedaryl, arylalkyl, and substituted arylalkyl; R² and R³ are the same ordifferent and, at each occurrence, independently hydrogen, alkyl,substituted alkyl, alkylcarboxyl or substituted alkylcarboxyl; or R² andR³ together and the carbon to which they are attached form a substitutedor unsubstituted five membered lactone; R⁴ is hydrogen, alkyl orsubstituted alkyl; and R⁵ is hydrogen, alkyl or substituted alkyl; 2)administering said compound to said subject.

In further embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted compound ofthe following formula:

or salt thereof functioning to decrease choroidal neovascularizationwherein, R¹ is hydrogen, alkyl, substituted alkyl, aryl, substitutedaryl, arylalkyl, and substituted arylalkyl; R² and R³ are the same ordifferent and, at each occurrence, independently hydrogen, alkyl,substituted alkyl, alkylcarboxyl or substituted alkylcarboxyl; or R² andR³ together and the carbon to which they are attached form a substitutedor unsubstituted five membered lactone; R⁴ is hydrogen, alkyl orsubstituted alkyl; and R⁵ is hydrogen, alkyl or substituted alkyl; 2)administering said compound to said subject.

In additional embodiments, the invention relates to a method of treatingor preventing macular degeneration comprising: I) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a compound having the following structure:

or salt thereof wherein, R¹ and R² are the same or different and, ateach occurrence, independently hydrogen or aryl; R³ and R⁴ are the sameor different and, at each occurrence, independently hydrogen, halogen,or alkyl; X is ═O or two hydrogens each independently bonded to thecarbon by a single bond and 2) administering said compound to saidsubject. In further embodiments, said macular degeneration isage-related. In further embodiments, said subject is a human. In furtherembodiments, said administration is topically to the eye. In furtherembodiments, said salt is a hydrochloride salt. In further embodiments,said composition is a liquid solution.

In additional embodiments, the invention relates to a method of treatingor preventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a compound having the following structure:

or salt thereof wherein, R¹ and R² are the same or different and, ateach occurrence, independently hydrogen or aryl; R³ and R⁴ are the sameor different and, at each occurrence, independently hydrogen, halogen,or alkyl; X is ═O or two hydrogens each independently bonded to thecarbon by a single bond and 2) administering said compound to saidsubject. In further embodiments, said macular degeneration isage-related. In further embodiments, said subject is a human. In furtherembodiments, said administration is topically to the eye. In furtherembodiments, said salt is a hydrochloride salt. In further embodiments,said composition is a liquid solution.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted compound having the followingstructure:

or salt thereof wherein, R¹ and R² are the same or different and, ateach occurrence, independently hydrogen, aryl or substituted aryl; R³and R⁴ are the same or different and, at each occurrence, independentlyhydrogen, halogen, alkyl or substituted alkyl; X is ═O or two hydrogenseach independently bonded to the carbon by a single bond; and 2)administering said compound to said subject.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted compoundhaving the following structure:

or salt thereof wherein, R¹ and R² are the same or different and, ateach occurrence, independently hydrogen, aryl or substituted aryl; R³and R⁴ are the same or different and, at each occurrence, independentlyhydrogen, halogen, alkyl or substituted alkyl; X is ═O or two hydrogenseach independently bonded to the carbon by a single bond; and 2)administering said compound to said subject.

In additional embodiments, the invention relates to a method of treatingor preventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising flavon and 2) administering said compound to said subject. Infurther embodiments, said macular degeneration is age-related. Infurther embodiments, said subject is a human. In further embodiments,said administration is topically to the eye. In further embodiments,said composition is a liquid solution.

In additional embodiments, the invention relates to a method of treatingor preventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising flavon and 2) administering said compoundto said subject. In further embodiments, said macular degeneration isage-related. In further embodiments, said subject is a human. In furtherembodiments, said administration is topically to the eye. In furtherembodiments, said composition is a liquid solution.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted compound of the followingformula:

functioning to decrease choroidal neovascularization and 2)administering said compound to said subject. In further embodiments,said substituted compound is quercetin, apigenin or puerarin. In furtherembodiments, said subject is a human. In further embodiments, saidadministration is topically to the eye. In further embodiments, saidcomposition is a liquid solution.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted compound ofthe following formula:

functioning to decrease choroidal neovascularization and 2)administering said compound to said subject. In further embodiments,said substituted compound is quercetin, apigenin or puerarin. In furtherembodiments, said subject is a human. In further embodiments, saidadministration is topically to the eye. In further embodiments, saidcomposition is a liquid solution.

In additional embodiments, the invention relates to a method of treatingor preventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and a compositioncomprising prednisolone and salts thereof and 2) administering saidcompound to said subject. In further embodiments, said maculardegeneration is age-related. In further embodiments, said subject is ahuman. In further embodiments, said administration is topically to theeye. In further embodiments, said composition is a liquid solution.

In additional embodiments, the invention relates to a method of treatingor preventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising prednisolone and salts thereof and 2)administering said compound to said subject. In further embodiments,said macular degeneration is age-related. In further embodiments, saidsubject is a human. In further embodiments, said administration istopically to the eye. In further embodiments, said composition is aliquid solution.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted compound of the followingformula:

and salts thereof wherein, R¹ is hydrogen, phosphate, alkylcarbonyl, orsuccinyl and R² is hydrogen or alkyl, and X is ═O or one hydrogen andone hydroxyl each independently bonded to the carbon by a single bond;functioning to decrease choroidal neovascularization and 2)administering said compound to said subject. In further embodiments,said salt is a sodium salt. In further embodiments, said compound isprednisolone, prednisolone acetate, prednisolone sodium phosphate,prednisolone tebutate, prednisone, methylprednisolone,methylprednisolone acetate, or methylprednisolone sodium succinate.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted compound ofthe following formula:

and salts thereof wherein, R¹ is hydrogen, phosphate, alkylcarbonyl, orsuccinyl and R² is hydrogen or alkyl, and X is ═O or one hydrogen andone hydroxyl each independently bonded to the carbon by a single bond;functioning to decrease choroidal neovascularization and 2)administering said compound to said subject. In further embodiments,said salt is a sodium salt. In further embodiments, said compound isprednisolone, prednisolone acetate, prednisolone sodium phosphate,prednisolone tebutate, prednisone, methylprednisolone,methylprednisolone acetate, or methylprednisolone sodium succinate.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a compound having the formula:

wherein R¹ is hydrogen, nitro, alkyl, or —(C═O)R⁴, R² is hydrogen,nitro, alkyl or halogen, R³ is hydrogen or alky, and R⁴ is hydroxyl,—NHOMe, or

and; 2) administering said compound to said subject. In furtherembodiments, said subject is a human. In further embodiments, saidadministration is topically to the eye.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: I) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a compound having the formula:

wherein R¹ is hydrogen, nitro, alkyl, or —(C═O)R⁴, R² is hydrogen,nitro, alkyl or halogen, R³ is hydrogen or alky, and R⁴ is hydroxyl,—NHOMe, or

and; 2) administering said compound to said subject. In furtherembodiments, said subject is a human. In further embodiments, saidadministration is topically to the eye.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted a compound having the formula:

wherein R¹ is hydrogen, nitro, alkyl, or —(C═O)R⁴, R² is hydrogen,nitro, alkyl or halogen, R³ is hydrogen or alky, and R⁴ is hydroxyl,—NHOMe, or

or salt thereof functioning to decrease choroidal neovascularization;and 2) administering said compound to said subject.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted a compoundhaving the formula:

wherein R¹ is hydrogen, nitro, alkyl, or —(C═O)R⁴, R² is hydrogen,nitro, alkyl or halogen, R³ is hydrogen or alky, and R⁴ is hydroxyl,—NHOMe, or

or salt thereof functioning to decrease choroidal neovascularization;and 2) administering said compound to said subject.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted a compound having the formula:

wherein R¹ is hydrogen, nitro, alkyl, or —(C—O)R⁴, B is N or C—R², R² ishydrogen, nitro, alkyl or halogen, R³ is hydrogen, nitro, —CO₂H, oralky, R⁴ is hydroxyl, —NHOMe, or

A is C—H or N, and R⁵ is hydrogen, —CH₂CO₂H, or nitro. In a preferredembodiment, R³ is nitro and R⁵ is hydrogen. In another preferredembodiment, B is C—R² wherein R² is nitro and R⁵ is hydrogen. In anotherpreferred embodiments, R¹ is nitro and R⁵ is hydrogen and R³ is —CO₂H orsalt thereof functioning to decrease choroidal neovascularization; and2) administering said compound to said subject.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted a compoundhaving the formula:

wherein R¹ is hydrogen, nitro, alkyl, or —(C═O)R⁴, B is N or C—R², R² ishydrogen, nitro, alkyl or halogen, R³ is hydrogen, nitro, —CO₂H, oralky, R⁴ is hydroxyl, —NHOMe, or

A is C—H or N, and R⁵ is hydrogen, —CH₂CO₂H, or nitro. In a preferredembodiment, R³ is nitro and R³ is hydrogen. In another preferredembodiment, B is C—R² wherein R² is nitro and R⁵ is hydrogen. In anotherpreferred embodiments, R¹ is nitro and R⁵ is hydrogen and R³ is —CO₂H orsalt thereof functioning to decrease choroidal neovascularization; and2) administering said compound to said subject.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted compound having the formula:

and derivatives thereof and 2) administering said compound to saidsubject. In further embodiments, said macular degeneration isage-related. In further embodiments, said subject is a human. In furtherembodiments, said administration is topically to the eye. In furtherembodiments, said composition is a liquid solution. In furtherembodiments, said subject is diagnosed with macular degeneration.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted compoundhaving the formula:

and derivatives thereof and 2) administering said compound to saidsubject. In further embodiments, said macular degeneration isage-related. In further embodiments, said subject is a human. In furtherembodiments, said administration is topically to the eye. In furtherembodiments, said composition is a liquid solution. In furtherembodiments, said subject is diagnosed with macular degeneration.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectdiagnosed with or at risk for macular degeneration and ii) a compositioncomprising a substituted or unsubstituted compound having the formula:

and derivatives thereof functioning to decrease choroidalneovascularization and 2) administering said compound to said subject.In further embodiments, said macular degeneration is age-related. Infurther embodiments, said subject is a human. In further embodiments,said administration is topically to the eye. In further embodiments,said composition is a liquid solution. In further embodiments, saidsubject is diagnosed with macular degeneration.

In some embodiments, the invention relates to a method of treating orpreventing macular degeneration comprising: 1) providing i) a subjectthat exhibits a symptom of macular degeneration, and in furtherembodiments said administering causes a reduction in said symptom andii) a composition comprising a substituted or unsubstituted compoundhaving the formula:

and derivatives thereof functioning to decrease choroidalneovascularization and 2) administering said compound to said subject.In further embodiments, said macular degeneration is age-related. Infurther embodiments, said subject is a human. In further embodiments,said administration is topically to the eye. In further embodiments,said composition is a liquid solution. In further embodiments, saidsubject is diagnosed with macular degeneration.

In additional embodiments, the invention relates to a method ofmanaging, prevention, and/or treating age-related macular degenerationcomprising: 1) providing i) a subject and ii) a composition comprising acompound functioning to decrease choroidal neovascularization and 2)administering said compound to said subject. In further embodiments,said compound is an interleukin-1 (IL-1) blockers preferably CK-17(5-bromo-5-methyl-3-phenyl-2-phenylimino-1,3-thiazinan-4-one), CK-112,CK-113, CK-115, CK-116, and CK-117. In further embodiments, saidcompound is a substituted or unsubstituted compound or derivative of thefollowing structure:

In further embodiments, said compound has the following structure:

wherein, R¹ is hydrogen, alkyl, substituted alkyl, aryl, substitutedaryl, arylalkyl, and substituted arylalkyl; R² and R³ are the same ordifferent and, at each occurrence, independently hydrogen, alkyl,substituted alkyl, alkylcarboxyl or substituted alkylcarboxyl; R⁴ ishydrogen or alkyl; and R⁵ is alkyl.

In further embodiments, said compound is selected from the groupconsisting hydralazine i.e., phthalazin-1-ylhydrazine, quanabenz, i.e.,(2-[(2,6-dichlorophenyl)methylideneamino]guanidine, D-timolol, i.e.,(2R)-1-[(4-morpholin-4-yl-1,2,5-thiadiazol-3-yl)oxy]-3-(tert-butylamino)propan-2-ol,apigenin i.e., 4,5-dihydroxy-2-(4-hydroxyphenyl)chromen-7-one,naringenin, i.e., 5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one,quercetin, i.e., 2-(3,4-dihydroxyphenyl)-3,4,5-trihydroxy-chromen-7-one,flavon, i.e., 3-hydroxy-2-phenyl-chromen-4-one, DN-6, i.e.,N-methoxy-1-nitro-1H-pyrazole-3-carboxamide, DN-7, i.e.,1-nitro-1H-pyrazole-3-carboxylic acid, and DN-13, i.e.,4-bromo-1-nitro-1H-pyrazole or substituted compounds and combinationsthereof.

In some embodiments the invention relates to the use of a compoundfunctioning to decrease choroidal neovascularization for the manufactureof a medicament for the treatment of macular degeneration, preferablyage-related macular degeneration.

In some embodiments, the invention relates to a compound or derivativeof a compound herein functioning as an interleukin-1 blocker used in thetreatment or prevention of eye diseases. In further embodiments, saidinterleutkin-1 blocker is selected from the group consisting of CK-17,CK-112, CK-113, CK-115, CK-116, and CK-117. In further embodiments, saidcompound is prednisolone. In further embodiments, said compound is asubstituted or unsubstituted compound or derivative of the followingstructure:

In further embodiments, said compound has the following structure:

wherein, R¹ is hydrogen, alkyl, substituted alkyl, aryl, substitutedaryl, arylalkyl, and substituted arylalkyl; R² and R³ are the same ordifferent and, at each occurrence, independently hydrogen, alkyl,substituted alkyl, alkylcarboxyl or substituted alkylcarboxyl; R⁴ ishydrogen or alkyl; and R⁵ is alkyl.

In some embodiments, the invention relates to the use of substituted orunsubstituted compounds disclosed herein or derivatives thereof for thetreatment of age-related macular degeneration.

In some embodiments, the present invention provides methods of treatingor preventing eye disease comprising administering an IL-1 blocker to apatient with eye-disease such that at least one symptom of the eyedisease is reduced or eliminated. In further embodiments, the eyedisease is age-related macular degeneration. In some embodiments, theadministering is performed via the eye. In certain embodiments, theadministering is performed orally or systemically. In furtherembodiment, the subject is diagnosed or at risk for maculardegeneration.

In particular embodiments, the present invention provides compositionscomprising; i) an IL-1 blocker, and ii) an ophthalmic solution.

In other embodiments, the present invention provides systems comprising:a) a composition comprising an IL-1 blocker and an ophthalmic solution;and b) an eye-dropper. In particular embodiments, the composition islocated in the eye-dropper.

In certain embodiments, the IL-blocker is specific for IL-1 alpha. Inother embodiments, the IL-blocker is specific for IL-1 beta. In furtherembodiments, the IL-1 blocker is selected from the group consisting ofCK-17, CK-112, CK-113, CK-115, CK-116, CK-117, CK-101A, CK-103A, CK-119,CK-120, and CK-122, and similar compounds. In other embodiments, theIL-1 blocker is selected from the group consisting of: IL-1 siRNAsequences configured to reduce the expression of IL-1 proteins, a vectorconfigured to express IL-1 siRNA sequences, anti-IL-1 antibodies orfragments, anti-IL1 antisense sequences, and vectors configured toexpress anti-IL1 antisense sequences.

In some embodiments, the invention relates to a method of treating orpreventing age-related macular degeneration comprising: 1) providing i)a subject diagnosed or at risk for macular degeneration and ii) acomposition comprising a compound functioning to decrease choroidalneovascularization and 2) administering said compound to said subject.In further embodiments, said compound is an interleukin-1 blocker. Infurther embodiments, said compound is selected from the group consistingof CK-17, CK-112, CK-113, CK-115, CK-116, CK-117, CK-101A, CK-103A,CK-119, CK-120, and CK-122. In further embodiments, said compound isprednisolone.

In additional embodiments, the invention relates to the use of acompound functioning to decrease choroidal neovascularization for themanufacture of a medicament for the treatment of macular degeneration.Preferably age-related macular degeneration.

In additional embodiments, the invention relates to the use of compounddisclosed herein for the manufacture of a medicament for the treatmentof age-related macular degeneration. Preferably age-related maculardegeneration.

In additional embodiments, the invention relates to the use of asubstituted compound disclosed herein functioning to decrease choroidalneovascularization for the manufacture of a medicament for the treatmentof age-related macular degeneration. In further embodiments, saidcompound is an interleukin-1 blocker. In further embodiments, saidcompound is selected from the group consisting of CK-17, CK-112, CK-113,CK-I 15, CK-116, CK-117, CK-10I A, CK-103A, CK-119, CK-120, and CK-122.In further embodiments, said compound is prednisolone, tetrandrine orosthole or derivative or substituted compound thereof. In furtherembodiments, said compound is a substituted or unsubstituted compound orderivative of the following structure:

In additional embodiments, the invention relates to the use of acompounds disclosed herein functioning to inhibit IL-1 induced uveitisfor the manufacture of a medicament for the treatment of arthritis,preferably rheumatoid arthritis. In further embodiment a subject showssymptoms of arthritis before administration. In further embodiments,said compound is an interleukin-1 blocker. In further embodiments, saidcompound is selected from the group consisting of CK-17, CK-112, CK-113,CK-115, CK-116, CK-117, CK-101A, CK-103A, CK-119, CK-120, and CK-122. Infurther embodiments, said compound is prednisolone, tetrandrine orosthole or derivative or substituted compound thereof.

The compositions comprising the active compounds of the presentinvention may include nutritional/dietary supplements and bulk-drugcompositions useful in the manufacture of pharmaceutical compositions(e.g., impure or non-sterile compositions) and pharmaceuticalcompositions (i.e., compositions that are suitable for administration toa subject) that can be used in the preparation of unit dosage forms.Such compositions optionally comprise a prophylactically ortherapeutically effective amount of a prophylactic and/or therapeuticagent disclosed herein or a combination of those agents and apharmaceutically acceptable carrier. Preferably, compositions of theinvention comprise a prophylactically or therapeutically effectiveamount of the active compound and another therapeutic or prophylacticagent, and a pharmaceutically acceptable carrier. These compositions maycontain between 0.1-99% of the active ingredient.

In certain embodiments, the therapeutic compounds (e.g., IL-1 blockers)are in an ophthalmic solution (e.g. such that the blockers can beadministered directly to the eye of the patient via an eye dropper).Preferably, the osmotic value of the solution is about 0.9% sodiumchloride (e.g. 0.6% to about 1%). It is also preferred that theophthalmic solution have a pH of about 7.4 (e.g. 6.6 to 7.8, preferably7.0 to 7.4). It is also preferred that the ophthalmic solution bebuffered to prevent wide changes in the pH.

BRIEF DESCRIPTION OF THE DRAWINGS

For a more complete understanding of the features and advantages of thepresent invention, reference is now made to the detailed description ofthe invention along with the accompanying figures.

FIG. 1 is a fluorescein angiograph that shows the control formation ofchoroidal neovascularization (CNV) by using a laser treatment.

FIG. 2 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 3 mg/kg 10 mg/kghydralazine.

FIG. 3 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 20 mg/kg guanabenz.

FIG. 4 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 3 mg/kg prednisolone.

FIG. 5 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 30 mg/kg CK-17.

FIG. 6 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 10 mg/kg CK-112.

FIG. 7 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 30 mg/kg CK-112.

FIG. 8 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 30 mg/kg CK-113.

FIG. 9 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 10 mg/kg CK-115.

FIG. 10 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 10 mg/kg CK-116.

FIG. 11 is a fluorescein angiograph that shows the inhibition of CNVformation by intraperitonal administration of 10 mg/kg CK-117.

FIG. 12 shows data of blockade of IL-1 Induced Uveitis by Synthetic IL-1Blockers: *, significantly different from corresponding controls atp<0.05 with N=6 eyes and Mean±SEM: **, 10 mg/kg, ip, t.i.d.

FIG. 13 show data of blockade of IL-1 Induced Uveitis by NaturalProducts: * significantly different from corresponding controls atp<0.15 with N=6 eyes and Mean±SEM.

FIG. 14 shows the delay of Trabeculaectomy Failure Caused byInflammation with Prednisolone and IL-1 Blockers *, significantly largerthan control at p<0.05 with N=6 eyes and Mean±SEM: ** 10 mg/injection atsubTenons

FIG. 15 shows data on the affects of aspirin and CK-17 on carrageenininduced inflammation: a, significantly different from controls at p<0.05with n=8 and (Mean±SD): b, significantly different from aspirin atp<0.05 with n=8 and (Mean±SD)

FIG. 16 shows data on rabbit irritation responses by 0.1% CK-17 afterthe eye-drop instillation. Scores are means of 6 eyes: Cornea=degree ofopacity; Iris=degree of iritis; Conjunctiva=redness, chemosis, anddischarge; Total Cornea, iris, and conjunctiva together; R=right eye(test); L=Left eye (control).

FIG. 17 shows the chemical structures of CK-17, CK-112, CK-113, CK-115,CK-116, and CK-117.

FIG. 18 shows the chemical structures of CK-101A, CK-103A, CK-119,CK-120, and CK-122.

FIG. 19 shows the chemical structures of DN-4 through DN-15.

FIG. 20 shows the chemical structures of DC-1 through DC-17.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates in general to therapeutic compositions andmethods of use. In preferred embodiments, the invention relates to thefield of eye health. In some embodiments, the invention relates to theprevention and treatment of macular degeneration by administeringcompounds disclosed herein. In some embodiments, the invention relatesto compositions and methods of improving vision.

In some embodiments, the invention relates to the prevention andtreatment of age-related macular degeneration by administering compoundsdisclosed herein. In some embodiments, the invention relates tocompositions and methods of improving health including dietarysupplements that prevent of choroid ischemic trauma leading toage-related macular degeneration.

Numerous methods have been attempted to treat age-related maculardegeneration without success. They include laser photocoagulation forchoroidal neovascularization, radiation treatment, transpupillarythermotherapy of subfoveal occult choroidal neovascularization,submacular surgery, limited macular translocation, adjuncts in surgery,argon laser to drusen, infrared diode laser photocoagulation.

Pharmacological treatments have been tried but with very limitedsuccess. For example, photodynamic therapy with verteporfin, visudyne,and BPD-MA has been shown to be beneficial for some wet-AMD patients(15%) but not for dry-AMD patients (85%). More recently, newer agentssuch as vascular endothelial growth factor (VEGF) receptor kinaseinhibitors, anti-VEGF antibodies, pigment epithelium-derived factor(PEDF), and angiostatin have been tried to prevent the CNV formation atthe very late stage of AMD. They are still in the experimental stage andnone have been shown to be efficacious in human patients. Thus, there isa need to identify agents that can be used to manage, prevent, and/ortreat age-related macular degeneration.

To facilitate the understanding of this invention, a number of terms aredefined below. Terms defined herein have meanings as commonly understoodby a person of ordinary skill in the areas relevant to the presentinvention. Terms such as “a”, “an” and “the” are not intended to referto only a singular entity, but include the general class of which aspecific example may be used for illustration. The terminology herein isused to describe specific embodiments of the invention, but their usagedoes not delimit the invention, except as outlined in the claims.

As used herein an “eye disease” means any variety of diseases,impairments, or defects that cause, vision loss, blurred or decreasedcentral close-up and distance vision, blind spots, objects to appear adifferent color or shape, neuro-ophthalmic manifestations of vasculareye diseases, including ischemic optic neuropathy, anterior ischemicoptic neuropathy, retinal artery occlusion, asymptomatic retinal emboli,asymptomatic retinal embolus or ischemia of retinal tissue, retinaledema, amaurosis fugax, reduction in visual field, occlusion of ocularvessels, stagnation of blood flow within the arteriole, cataracts,glaucoma, proptosis, eyelid retraction, restrictive myopathy, diplopia(double vision), compressive optic neuropathy, and/or exposurekeratopathy. In a preferred embodiment, the eye disease is maculardegeneration or diabetic eye disease. It is not intended that thepresent invention be limited to treating any particular underlyingdisease resulting in vision defects or impairments.

As used herein, “macular degeneration” means any condition that causespart of the macula to deteriorate. This degeneration may be partial ortotal, and it is not intended to be limited to advance stages of thedisease: thus, is intended to include a subject that is diagnosed withdrusen even thought the subject does not have any symptoms of impairedvision. A symptom of macular degeneration is a change in central vision.The patient may notice blurred central vision or a blank spot on thepage when reading. The patient may notice visual distortion such asbending of straight lines. Images may appear smaller. Some patientsnotice a change in color perception and some experience abnormal lightsensations. These symptoms may come on suddenly and become progressivelymore troublesome. Sudden onset of symptoms, particularly visiondistortion, is an indication for immediate evaluation by anophthalmologist.

As used herein a diagnosis of macular degeneration means any analysis ofmacular changes or function in a subject. It is not intended to belimited to any particular method. For example, an eye examiner, e.g.,doctor, may dilate the pupil with eye drops and examine the interior ofthe eye, looking at the retina for the presence of yellow bumps ofdrusen, eye lesions, or for gross changes in the macula such asthinning. The eye examiner may also administer a visual field test,looking for blank spots in the central vision. The examiner may call forfluorescein angiography (intravenous injection of fluorescent dyefollowed by visual examination and photography of the back of the eye)to determine if blood vessels in the retina are leaking.

Some risk factors for having macular degeneration include, age, smoking,and a diet that is rich in saturated fat. Others may be at risk formacular degeneration because of genetic heritage or environmentalexposure. In preferred embodiments, the invention relates to treating orprevention of age-related macular degeneration, preferably prophylacticprevention and treatment.

Age-related macular degeneration may be characterized as a dry(atrophic) or wet (exudative) form. Multiple, small, round, yellow-whitespots called drusen are identifiers for the dry type. The spots aretypically located in the back of the eye at the level of the outerretina. Subjects with these spots may have excellent vision and nosymptoms. Most subjects with age-related macular degeneration begin withthe dry form. In the wet form, newly created abnormal blood vessels growunder the center of the retina. These blood vessels leak, bleed, andscar the retina, distorting vision or destroying central vision. Visiondistortion may starts in one eye and may affect the other eye later.

A diabetic (Type I or Type II) patient is at risk for maculardegeneration. Diabetic macular degeneration is the deterioration of themacula due to diabetes. Cystoid macular degeneration is the loss ofvision in the macula due to fluid-filled areas (cysts) in the macularregion. This may be a result of other disorders, inflammation, or highmyopia.

As used herein, a compound “functioning to decrease choroidalneovascularization” means that a statistically significant reduction ofchoroidal neovascularization can be measured, e.g. by fluoresceinangiography, after some period of time of administering said compound toa mammalian after physical disruption of the eye's Bruch's membrane,e.g., via a laser. A detailed description of these methods foridentifying compounds functioning to decrease choroidalneovascularization are describe herein.

“Isomers” means any of two or more substances that are composed of thesame elements in the same proportions but differ in the threedimensional arrangement of atoms including enantiomeric (i.e., mirrorimages) and diastereomeric isomers.

“Timolol compound” or molecules, and the like means substituted orunsubstituted compounds of the following formula:

As used herein, the term “timolol component” refers that part of acomposition that contains all of timolol molecules in a givencomposition, including all conformational and stereomeric forms. Inpreferred embodiments, a given compound (e.g. designated by a structure)makes up a large percentage (e.g. by number of molecules and/or byweight) of the timolol component. For example, a given timololderivative may be present in an aqueous composition at a level where 70%of all the timolol components are of that given compound, e.g.D-Timolol, while most of the composition itself is composed of water.

The term “salts”, as used herein, refers to any salt that complexes withidentified compounds contained herein while retaining a desiredfunction, e.g., biological activity. Examples of such salts include, butare not limited to, acid addition salts formed with inorganic acids(e.g. hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoricacid, nitric acid, and the like), and salts formed with organic acidssuch as, but not limited to, acetic acid, oxalic acid, tartaric acid,succinic acid, malic acid, fumaric acid, maleic acid, ascorbic acid,benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic,acid, naphthalene sulfonic acid, naphthalene disulfonic acid, andpolygalacturonic acid. Salt compounds can also be administered aspharmaceutically acceptable quaternary salts known by a person skilledin the art, which specifically include the quaternary ammonium salts ofthe formula —NR,R′,R″⁺Z⁻, wherein R, R′, R″ is independently hydrogen,alkyl, or benzyl, and Z is a counter ion, including, but not limited to,chloride, bromide, iodide, alkoxide, toluencsulfonate, methylsulfonate,sulfonate, phosphate, or carboxylate (such as benzoate, succinate,acetate, glycolate, maleate, malate, fumarate, citrate, tartrate,ascorbate, cinnamoate, mandeloate, and diphenylacetate).

“Adverse drug reaction” means any response to a drug that is noxious andunintended and occurs in doses for prophylaxis, diagnosis, or therapyincluding side effects, toxicity, hypersensitivity, drug interactions,complications, or other idiosyncrasy. Side effects are often adversesymptom produced by a therapeutic serum level of drug produced by itspharmacological effect on unintended organ systems (e.g., blurred visionfrom anticholinergic antihistamine). A toxic side effect is an adversesymptom or other effect produced by an excessive or prolonged chemicalexposure to a drug (e.g., digitalis toxicity, liver toxicity).Hypersensitivities are immune-mediated adverse reactions (e.g.,anaphylaxis, allergy). Drug interactions are adverse effects arisingfrom interactions with other drugs, foods or disease states (e.g.,warfarin and erythromycin, cisapride and grapefruit, loperamide andClostridium difficile colitis). Complications are diseases caused by adrug (e.g., NSAID-induced gastric ulcer, estrogen-induced thrombosis).The adverse drug reaction may be mediated by known or unknown mechanisms(e.g., Agranulocytosis associated with chloramphenicol or clozapine).Such adverse drug reaction can be determined by subject observation,assay or animal model well-known in the art.

“Alkyl” means a straight chain or branched, noncyclic or cyclic,unsaturated or saturated aliphatic hydrocarbon containing from 1 to 10carbon atoms, while the term “lower alkyl” has the same meaning as alkylbut contains from 1 to 6 carbon atoms. The term “higher alkyl” has thesame meaning as alkyl but contains from 2 to 10 carbon atoms.Representative saturated straight chain alkyls include methyl, ethyl,n-propyl, n-butyl, n-pentyl, n-hexyl, n-septyl, n-octyl, n-nonyl, andthe like; while saturated branched alkyls include isopropyl, sec-butyl,isobutyl, tert-butyl, isopentyl, and the like. Representative saturatedcyclic alkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,and the like; while unsaturated cyclic alkyls include cyclopentenyl andcyclohexenyl, and the like. Cyclic alkyls are also referred to herein asa “homocycles” or “homocyclic rings.” Unsaturated alkyls contain atleast one double or triple bond between adjacent carbon atoms (referredto as an “alkenyl” or “alkynyl”, respectively). Representative straightchain and branched alkenyls include ethylenyl, propylenyl, 1-butenyl,2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl,2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, and the like; whilerepresentative straight chain and branched alkynyls include acetylenyl,propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl,3-methyl-1-butynyl, and the like.

“Alkylamino” and “dialkylamino” mean one or two alkyl moiety attachedthrough a nitrogen bridge (i.e., —N-alkyl) such as methylamino,ethylamino, dimethylamino, diethylamino, and the like.

“Alkylcarboxyl” means an alkyl moiety attached through a carboxyl group(i.e., CO₂Alkyl).

“Alkylcarboxyl” means an alkyl moiety attached through a carbonyl group(i.e., —(C═O)Alkyl). A “succinyl” is an ethylcarbonyl substituted with acarboxyl group on the second carbon (i.e., —(C═O)CH₂CH₂CO₂H).

“Alkylthiol” means an alkyl moiety attached through a sulfur bridge(i.e., —S-alkyl).

“Alkyloxy” means an alkyl moiety attached through an oxygen bridge(i.e., —O-alkyl) such as methoxy, ethoxy, and the like.

“Alkoxide” means an alkyl moiety attached to a negatively charged oxygenatom (i.e., —Oalkyl) such as methoxide or ethoxide.

Within the context of certain embodiment, an “amine” group means —NH₂,and “ammonia” means the gas NH₃.

“Aryl” means an aromatic carbocyclic moiety such as phenyl or naphthyl.

“Arylalkyl” means an aryl moiety attached through an alkyl bridge (e.g.,—CH₂-Phenyl).

“Aryloxy” means an aryl moiety attached through an oxygen bridge (i.e.,—O-aryl).

“Arylthio” means an alkyl moiety attached through a sulfur bridge (i.e.,—S-aryl).

“Heteroaryl” means an aromatic heterocycle ring of 5- to 10 members andhaving at least one heteroatom selected from nitrogen, oxygen andsulfur, and containing at least 1 carbon atom, including both mono- andbicyclic ring systems. Representative heteroaryls are furyl,benzofuranyl, thiophenyl, benzothiophenyl, pyrrolyl, indolyl,isoindolyl, azaindolyl, pyridyl, quinolinyl, isoquinolinyl, oxazolyl,isooxazolyl, benzoxazolyl, pyrazolyl, imidazolyl, benzimidazolyl,thiazolyl, benzothiazolyl, isothiazolyl, pyridazinyl, pyrimidinyl,pyrazinyl, triazinyl, cinnolinyl, phthalazinyl, and quinazolinyl.

“Heteroarylalkyl” means an alkyl having at least one alkyl hydrogen atomreplaced with a heteroaryl moiety, such as —CH₂pyridinyl,—CH₂pyrimidinyl, and the like.

“Heterocycle” (also referred to herein as a “heterocyclic ring”) means a4- to 7-membered monocyclic, or 7- to 10-membered bicyclic, heterocyclicring which is either saturated, unsaturated, or aromatic, and whichcontains from 1 to 4 heteroatoms independently selected from nitrogen,oxygen and sulfur, and wherein the nitrogen and sulfur heteroatoms maybe optionally oxidized, and the nitrogen heteroatom may be optionallyquaternized, including bicyclic rings in which any of the aboveheterocycles are fused to a benzene ring. The heterocycle may beattached via any heteroatom or carbon atom. Heterocycles includeheteroaryls as defined above. Thus, in addition to the heteroarylslisted above, heterocycles also include morpholinyl, pyrrolidinonyl,pyrrolidinyl, piperidinyl, hydantoinyl, valerolactamyl, oxiranyl,oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl,tetrahydroprimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl,tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, andthe like.

“Heterocyclealkyl” means an alkyl having at least one alkyl hydrogenatom replaced with a heterocycle, such as —CH₂morpholinyl, and the like.

“Homocycle” (also referred to herein as “homocyclic ring”) means asaturated or unsaturated (but not aromatic) carbocyclic ring containingfrom 3-7 carbon atoms, such as cyclopropane, cyclobutane, cyclopentane,cyclohexane, cycloheptane, cyclohexene, and the like.

“Isomers” means any of two or more substances that are composed of thesame elements in the same proportions but differ in the threedimensional arrangement of atoms including enantiomeric (i.e., mirrorimages) and diastereomeric isomers.

The term “derivative” when used in relation to a chemical compoundrefers to a similar structure that upon application, e.g.,administration to a subject, is capable of providing, directly orindirectly, the function said chemical compound is disclosed to have(albeit the derivative may have increased or decreased function). Forexample, substituting one atom for another atom in a chemical compoundprovides a compound of similar structure, e.g., a carbon atom for anitrogen atom. The compound of similar structure may be capable offunctioning to decrease choroidal neovascularization. Certain claimedembodiments are intended to encompass minor changes in chemicalstructure provided that the derivative can decrease choroidalneovascularization.

The term “manage” when used in connection with a disease or conditionmeans to provide beneficial effects to a subject being administered witha prophylactic or therapeutic agent, which does not result in a cure ofthe disease. In certain embodiments, a subject is administered with oneor more prophylactic or therapeutic agents to manage a disease so as toprevent the progression or worsening of the disease.

As used herein, the terms “prevent” and “preventing” include theprevention of the recurrence, spread or onset. It is not intended thatthe present invention be limited to complete prevention. In someembodiments, the onset is delayed, or the severity of the disease isreduced.

“Subject” means any animal, preferably a human patient, livestock, ordomestic pet.

The term “substituted”, as used herein, means at least one hydrogen atomof a molecular arrangement is replaced with a substituent. In the caseof an oxo substituent (“═O”), two hydrogen atoms are replaced. Whensubstituted, one or more of the groups below are “substituents.”Substituents within the context of this invention include, but are notlimited to, halogen, hydroxy, oxo, cyano, nitro, amino, alkylamino,dialkylamino, alkyl, alkoxy, alkylthio, haloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocycle, and heterocyclealkyl, as wellas, —NR_(a)R_(b), —NR_(a)C(═O)R_(b), —NR_(a)C(═O)NR_(a)NR_(b),—NR_(a)C(═O)OR_(b)—NR_(a)SO₂R_(b), —C(═O)R_(a), C(═O)OR_(a),—C(═O)NR_(a)R_(b), —OC(═O)NR_(a)R_(b), —OR_(a), —SR_(a),—SOR_(a)—S(═O)₂R_(a), —OS(═O)₂R_(a), and —S(═O)₂OR_(a). In addition, theabove substituents may be further substituted with one or more of theabove substituents, such that the substituent comprises a substitutedalky, substituted aryl, substituted arylalkyl, substituted heterocycle,or substituted heterocyclealkyl. R_(a) and R_(b) in this context may bethe same or different and, independently, hydrogen, alkyl, haloalkyl,substituted alkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl orsubstituted heterocyclealkyl.

The term “unsubstituted”, as used herein, refers to any compound doesnot contain extra substituents attached to the compound. Anunsubstituted compound refers to the chemical makeup of the compoundwithout extra substituents, e.g., the compound does not containprotecting group(s). For example, unsubstituted proline is a prolineamino acid even though the amino group of proline may be considereddisubstituted with alkyl groups.

With regard to certain embodiments, a chemical structure may be drawnwith two lines between a first atom and substituent meaning that thereare two bonds, i.e., designate a double between the first atom and adefined substituent or it may designate two single bonds between thefirst atom and two defined substituent atoms.

As used herein, the terms “treat” and “treating” are not limited to thecase where the subject (e.g. patient) is cured and the disease iseradicated. Rather, the present invention also contemplates treatmentthat merely reduces symptoms, improves vision (to some degree) and/ordelays disease progression.

As used herein, the term “IL-1 blocker” refers to any compound orcomposition that is able to at least partially inhibit the biologicalactivity or expression of IL-1a and/or IL-1b in the eye of a patient. Incertain embodiments, IL-1 blockers bind to IL-1 proteins or RNAtranscripts. In other embodiments, IL-1 blockers competitively inhibitthe activity of IL-1 proteins. In further embodiments, the IL-1 blockersdestroy IL-1 proteins through cleavage. In some embodiments, the IL-1blockers cleave or bind to IL-1 mRNA transcripts. Examples of IL-1blockers include, but are not limited to, CK-17(5-bromo-5-methyl-3-phenyl-2-phenylimino-1,3-thiazinan-4-one), CK-112,CK-113, CK-115, CK-116, CK-117, IL-1 siRNA sequences and vectorsexpressing IL-1 siRNA sequences, IL-1 antisense sequences and vectorsexpressing IL-1 antisense sequences, anti-IL-1 antibodies and fragmentsthereof, including chimeric and preferably humanized or human anti-IL-1antibodies. Nucleic acid based IL-1 blockers, such as siRNA andantisense can be designed, for example, using the human IL-1a sequence(accession no. BC013142) or the human IL-1b sequence (accession numberBC008678) using techniques and software that are known in the art.Antibody based IL-1 blockers, such as anti-IL-1a or anti-IL-1bmonoclonal antibodies can be generated using the human IL-1a proteinsequence (accession no. CAG33695) or the human IL-1b protein sequence(accession no. CAG28607) using techniques known in the art.

The term “siRNAs” refers to short interfering RNAs. In some embodiments,siRNAs comprise a duplex, or double-stranded region, of about 18-25nucleotides long. Often siRNAs contain from about two to four unpairednucleotides at the 3′ end of each strand. At least one strand of theduplex or double-stranded region of a siRNA is substantially homologousto or substantially complementary to a target RNA molecule, such as IL-1mRNA transcripts. The strand complementary to a target RNA molecule isthe “antisense strand;” the strand homologous to the target RNA moleculeis the “sense strand,” and is also complementary to the siRNA antisensestrand. siRNAs may also contain additional sequences; non-limitingexamples of such sequences include linking sequences, or loops, as wellas stem and other folded structures. siRNAs appear to function as keyintermediaries in triggering RNA interference in invertebrates and invertebrates, and in triggering sequence-specific RNA degradation duringposttranscriptional gene silencing in plants.

The term “RNA interference” or “RNAi” refers to the silencing ordecreasing of gene expression by siRNAs. It is the process ofsequence-specific, post-transcriptional gene silencing in animals andplants, initiated by siRNA that is homologous in its duplex region tothe sequence of the silenced gene. The gene (e.g. IL-1a or IL-1b) may beendogenous or exogenous to the organism, present integrated into achromosome or present in a transfection vector that is not integratedinto the genome. The expression of the gene (e.g. IL-1a) is eithercompletely or partially inhibited. RNAi may also be considered toinhibit the function of a target IL-1 RNA; the function of the targetIL-1 RNA may be complete or partial.

Aging is a chronic process to cause degeneration of cells, tissues, andorgans, including choroidal blood vessels, retinal pigment epitheliumcells (RPEC) and Bruch's membrane of macular. Most notablyarteriosclerotic aging changes choroidal blood vessels, particularly themacular chorio-capillaris with a decrease in total capillary membraneand the blood flow. As a result, RPE starts to accumulate lipofuscin,alters cell shape, density, pigmentation, lysosomal activity andextracellular matrix formation. Gradually, Bruch's membrane showsthickening and decreased permeability, resulting with breakdown ofBruch's membrane which allows choroidal neovascularization (CNV) toappear.

Since CNV is formed because of vascular inflammation, others have triedto use VEGF receptor kinase inhibitors, anti-VEGF antibodies, PEDF andangiostatin to prevent the CNV formation and AMD worsening. The problemis these agents are used to remove only one of many inflammatory factorsand thus the efficacy is not obvious unless all these agents areadministered at the same time. While not necessary to understand orpractice the present invention, it is believed that a factor of vascularinflammation is caused by IL-1 and thus if IL-1 blockers are given, thevascular inflammation is decreased

The breakdown of Bruch's membrane enables the growth of CNV intosubfoveal space. Agents which stabilize Bruch's membrane also prevent itfrom breakdown and the penetration of CNV into the subfoveal space. Asit is believed that IL-1 causes pathological changes of Bruch'smembrane, IL-1 blockers stabilize it and prevent it from breaking down.Therefore, in certain embodiments, I1-1 blockers serve as double edgesword to prevent vascular inflammation and Bruch's membrane breakingdown so that CNV formation and spreading can be prevented and, in turn,so does AMD.

It is recognized that AMD preferably is treated at the earliest stagepossible to prevent disease progression. The earliest stage of AMDdevelopment is the malfunction of choroidal blood flow, resulting in adecrease of the blood flow of choriocapillaris. Chain reactions aretriggered that lead to RPE degenerations, Bruch's membrane breakdown,CNV formation, AMD and finally blindness. Therefore, specific drugs thatincrease the choroidal blood flow were found herein to be useful toprevent the AMD from developing and worsening. The agents discoveredinclude, but are not limited to: hypotensive agents (e.g., timolol,hydralazine guanaben); flavonoids (e.g., Apigenin, Naringenin Quercetin,Flavon); and N-nitro-pyrazoles and C-nitro-pyrazoles (e.g., DN-6, CN-7,DN-13, and DC-5).

Pharmaceutical Formulations

In a specific embodiment, the term “pharmaceutically acceptable” meansapproved by a regulatory agency of the Federal or a state government orlisted in the U.S. Pharmacopeia or other generally recognizedpharmacopeia for use in animals, and more particularly in humans. Theterm “carrier” refers to a diluent, adjuvant, excipient, or vehicle withwhich the active compound is administered. Such pharmaceutical vehiclescan be liquids, such as water and oils, including those of petroleum,animal, vegetable or synthetic origin, such as peanut oil, soybean oil,mineral oil, sesame oil and the like. The pharmaceutical vehicles can besaline, gum acacia, gelatin, starch paste, talc, keratin, colloidalsilica, urea, and the like. In addition, auxiliary, stabilizing,thickening, lubricating and coloring agents can be used. Whenadministered to a subject, the pharmaceutically acceptable vehicles arepreferably sterile. Water can be the vehicle when the active compound isadministered intravenously. Saline solutions and aqueous dextrose andglycerol solutions can also be employed as liquid vehicles, particularlyfor injectable solutions. Suitable pharmaceutical vehicles also includeexcipients such as starch, glucose, lactose, sucrose, gelatin, malt,rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate,talc, sodium chloride, dried skim milk, glycerol, propyleneglycol,water, ethanol and the like. The present compositions, if desired, canalso contain minor amounts of wetting or emulsifying agents, or pHbuffering agents.

The present compositions can take the form of solutions, suspensions,emulsion, tablets, pills, pellets, capsules, capsules containingliquids, powders, sustained-release formulations, suppositories,emulsions, aerosols, sprays, suspensions, or any other form suitable foruse. In one embodiment, the pharmaceutically acceptable vehicle is acapsule (see e.g., U.S. Pat. No. 5,698,155).

In a preferred embodiment, the active compound and optionally anothertherapeutic or prophylactic agent are formulated in accordance withroutine procedures as pharmaceutical compositions adapted forintravenous administration to human beings. Typically, the activecompound for intravenous administration are solutions in sterileisotonic aqueous buffer. Where necessary, the compositions can alsoinclude a solubilizing agent. Compositions for intravenousadministration can optionally include a local anesthetic such aslignocaine to ease pain at the site of the injection. Generally, theingredients are supplied either separately or mixed together in unitdosage form, for example, as a dry lyophilized powder or water freeconcentrate in a hermetically sealed container such as an ampoule orsachette indicating the quantity of active agent. Where the activecompound is to be administered by infusion, it can be dispensed, forexample, with an infusion bottle containing sterile pharmaceutical gradewater or saline. Where the active compound is administered by injection,an ampoule of sterile water for injection or saline can be provided sothat the ingredients can be mixed prior to administration.

Compositions for oral delivery can be in the form of tablets, lozenges,aqueous or oily suspensions, granules, powders, emulsions, capsules,syrups, or elixirs, for example. Orally administered compositions cancontain one or more optional agents, for example, sweetening agents suchas fructose, aspartame or saccharin; flavoring agents such aspeppermint, oil of wintergreen, or cherry; coloring agents; andpreserving agents, to provide a pharmaceutically palatable preparation.Moreover, where in tablet or pill form, the compositions can be coatedto delay disintegration and absorption in the gastrointestinal tractthereby providing a sustained action over an extended period of time.Selectively permeable membranes surrounding an osmotically activedriving compound are also suitable for an orally administered of theactive compound. in these later platforms, fluid from the environmentsurrounding the capsule is imbibed by the driving compound, which swellsto displace the agent or agent composition through an aperture. Thesedelivery platforms can provide an essentially zero order deliveryprofile as opposed to the spiked profiles of immediate releaseformulations. A time delay material such as glycerol monostearate orglycerol stearate can also be used. Oral compositions can includestandard vehicles such as mannitol, lactose, starch, magnesium stearate,sodium saccharine, cellulose, magnesium carbonate, and the like. Suchvehicles are preferably of pharmaceutical grade.

Further, the effect of the active compound can be delayed or prolongedby proper formulation. For example, a slowly soluble pellet of theactive compound can be prepared and incorporated in a tablet or capsule.The technique can be improved by making pellets of several differentdissolution rates and filling capsules with a mixture of the pellets.Tablets or capsules can be coated with a film that resists dissolutionfor a predictable period of time. Even the parenteral preparations canbe made long-acting, by dissolving or suspending the compound in oily oremulsified vehicles which allow it to disperse only slowly in the serum.

Compositions for use in accordance with the present invention can beformulated in conventional manner using one or more physiologicallyacceptable carriers or excipients.

Thus, the compound and optionally another therapeutic or prophylacticagent and their physiologically acceptable salts and solvates can beformulated into pharmaceutical compositions for administration byinhalation or insufflation (either through the mouth or the nose) ororal, parenteral or mucosol (such as buccal, vaginal, rectal,sublingual) administration. In some embodiments, the administration isoptical (e.g. eyes drops applied directly to the eye). In oneembodiment, local or systemic parenteral administration is used.

For oral administration, the compositions can take the form of, forexample, tablets or capsules prepared by conventional means withpharmaceutically acceptable excipients such as binding agents (e.g.,pregelatinised maize starch, polyvinylpyrrolidone or hydroxypropylmethylcellulose); fillers (e.g., lactose, microcrystalline cellulose orcalcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talcor silica); disintegrants (e.g., potato starch or sodium starchglycolate); or wetting agents (e.g., sodium lauryl sulphate). Thetablets can be coated by methods well known in the art. Liquidpreparations for oral administration can take the form of, for example,solutions, syrups or suspensions, or they can be presented as a dryproduct for constitution with water or other suitable vehicle beforeuse. Such liquid preparations can be prepared by conventional means withpharmaceutically acceptable additives such as suspending agents (e.g.,sorbitol syrup, cellulose derivatives or hydrogenated edible fats);emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles(e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetableoils); and preservatives (e.g., methyl or propyl-p-hydroxybenzoates orsorbic acid). The preparations can also contain buffer salts, flavoring,coloring and sweetening agents as appropriate.

Preparations for oral administration can be suitably formulated to givecontrolled release of the active compound.

For buccal administration the compositions can take the form of tabletsor lozenges formulated in conventional mariner.

For administration by inhalation, the compositions for use according tothe present invention are conveniently delivered in the form of anaerosol spray presentation from pressurized packs or a nebulizer, withthe use of a suitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol the dosage unitcan be determined by providing a valve to deliver a metered amount.Capsules and cartridges of e.g., gelatin for use in an inhaler orinsufflator can be formulated containing a powder mix of the compoundand a suitable powder base such as lactose or starch.

The compositions can be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion. Formulationsfor injection can be presented in unit dosage form, e.g., in ampoules orin multi-dose containers, with an added preservative. The pharmaceuticalcompositions can take such forms as suspensions, solutions or emulsionsin oily or aqueous vehicles, and can contain formulatory agents such assuspending, stabilizing and/or dispersing agents. Alternatively, theactive ingredient can be in powder Rhin for constitution with a suitablevehicle, e.g., sterile pyrogen-free water, before use.

In addition to the formulations described previously, the compositionscan also be formulated as a depot preparation. Such long actingformulations can be administered by implantation (for examplesubcutaneously or intramuscularly) or by intramuscular injection. Thus,for example, the pharmaceutical compositions can be formulated withsuitable polymeric or hydrophobic materials (for example as an emulsionin an acceptable oil) or ion exchange resins, or as sparingly solublederivatives, for example, as a sparingly soluble salt.

The compositions can, if desired, be presented in a pack or dispenserdevice that can contain one or more unit dosage forms containing theactive ingredient. The pack can for example comprise metal or plasticfoil, such as a blister pack. The pack or dispenser device can beaccompanied by instructions for administration.

In certain preferred embodiments, the pack or dispenser contains one ormore unit dosage forms containing no more than the recommended dosageformulation as determined in the Physician's Desk Reference (56^(th) ed.2002, herein incorporated by reference in its entirety).

Methods of administering the active compound and optionally anothertherapeutic or prophylactic agent include, but are not limited to,parenteral administration (e.g., intradermal, intramuscular,intraperitoneal, intravenous and subcutaneous), epidural, and mucosal(e.g., intranasal, rectal, vaginal, sublingual, buccal or oral routes).In a specific embodiment, the active compound and optionally anotherprophylactic or therapeutic agents are administered intramuscularly,intravenously, or subcutaneously. The active compound and optionallyanother prophylactic or therapeutic agent can also be administered byinfusion or bolus injection and can be administered together with otherbiologically active agents. Administration can be local or systemic. Theactive compound and optionally the prophylactic or therapeutic agent andtheir physiologically acceptable salts and solvates can also beadministered by inhalation or insufflation (either through the mouth orthe nose). In a preferred embodiment, local or systemic parenteraladministration is used.

In specific embodiments, it can be desirable to administer the activecompound locally to the area in need of treatment. This can be achieved,for example, and not by way of limitation, by local infusion duringsurgery or topical application, e.g., in conjunction with a wounddressing after surgery, by injection, by means of a catheter, by meansof a suppository, or by means of an implant, said implant being of aporous, non-porous, or gelatinous material, including membranes, such assilastic membranes, or fibers. In one embodiment, administration can beby direct injection at the site (or former site) of an atheroscleroticplaque tissue.

Pulmonary administration can also be employed, e.g., by use of aninhaler or nebulizer, and formulation with an aerosolizing agent, or viaperfusion in a fluorocarbon or synthetic pulmonary surfactant. Incertain embodiments, the active compound can be formulated as asuppository, with traditional binders and vehicles such astriglycerides.

In another embodiment, the active compound can be delivered in avesicle, in particular a liposome.

In yet another embodiment, the active compound can be delivered in acontrolled release system. In one embodiment, a pump can be used. Inanother embodiment, polymeric materials can be used.

The amount of the active compound that is effective in the treatment orprevention of age-related macular degeneration can be determined bystandard research techniques. For example, the dosage of the activecompound which will be effective in the treatment or prevention ofage-related macular degeneration can be determined by administering theactive compound to an animal in a model such as, e.g., the animal modelsknown to those skilled in the art. In addition, in vitro assays canoptionally be employed to help identify optimal dosage ranges.

Selection of a particular effective dose can be determined (e.g., viaclinical trials) by a skilled artisan based upon the consideration ofseveral factors which will be known to one skilled in the art. Suchfactors include the disease to be treated or prevented, the symptomsinvolved, the subject's body mass, the subject's immune status and otherfactors known by the skilled artisan.

The dose of the active compound to be administered to a subject, such asa human, is rather widely variable and can be subject to independentjudgment. It is often practical to administer the daily dose of theactive compound at various hours of the day. However, in any given case,the amount of the active compound administered will depend on suchfactors as the solubility of the active component, the formulation used,subject condition (such as weight), and/or the route of administration.

The general range of effective amounts of the active compound alone orin combination with another prophylactic or therapeutic agent(s) arefrom about 0.001 mg/day to about 1000 mg/day, more preferably from about0.001 mg/day to 750 mg/day, more preferably from about 0.001 mg/day to500 mg/day, more preferably from about 0.001 mg/day to 250 mg/day, morepreferably from about 0.001 mg/day to 100 mg/day, more preferably fromabout 0.001 mg/day to 75 mg/day, more preferably from about 0.001 mg/dayto 50 mg/day, more preferably from about 0.001 mg/day to 25 mg/day, morepreferably from about 0.001 mg/day to 10 mg/day, more preferably fromabout 0.001 mg/day to 1 mg/day. Of course, it is often practical toadminister the daily dose of compound in portions, at various hours ofthe day. However, in any given case, the amount of compound administeredwill depend on such factors as the solubility of the active component,the formulation used, subject condition (such as weight), and/or theroute of administration.

Choroidal Blood Flow

Naringenin is a flavon analog with potent effects to increase thechoroidal blood flow as can be seen in Table 1. The blood flow increasedrapidly at 30 min after drug administration. The effect peaked at 60 minafter drug administration and maintained the drug action for at least 2hrs. At the peak, the choroidal blood flow was increased more than 200%of the original blood flow.

Apigenin is a flavon analog. The drug actions lasted from 1 hr to 2 hrsafter administration (Table 1).

Puerarin is also a flavon analog similar to apigenin. Puerarin's actionstarted from 1 hr after administration and lasted beyond 3 hrs afteradministration (Table 1).

TABLE 1 Effects of Flavonoids on Choroidal Blood Flow Compounds (50 μl,1%) 30 min 60 min 120 min 180 min Vehicle 100^(a) 100^(b) 100^(c)100^(d) Naringenin   102.5 ± 22.0^(e)* 226.1 ± 55.0* 124.2 ± 74.5* 137.4± 712   Apingenin 9.7 ± 1.5 180.1 ± 42.2* 54.8 ± 6.7* −II 1 ± 2.3  Puerarin 30.9 ± 7.7  114.9 ± 21.5* 160.8 ± 70.7* 127.4 ± 65.5**Significantly higher than corresponding controls at 100% ^(a)5.9 ± 0.7μl/min/mg; ^(b)3.89 + 0.7 μl/min/mg; ^(c)2.2 ± 0.5 μl/min/mg; ^(d)1.71 +0.5 μl/min/mg; ^(e)mean ± SD with N = 6 for all except N = 10 forcontrols.

N-Nitropyrazoles, including DN-6, DN-7, DN-12, and DN-13 showed verystrong increase of choroidal blood flow and long duration of actionlasting beyond 3 hrs after drug administration (Table 2). DN-13 wasparticularly good as it acted rapidly to increase choroidal blood flowat 30 min after drug administration and the action lasted more than 3hrs after drug administration (Table 2). Among C-nitropyrazoles, DC-5showed the most promising actions with short latent period of less than30 min and long duration of action lasting longer than 3 hrs (Table 2).

TABLE 2 Effects N-nitro-azoles and C-nytropyrazoles on Choroidal BloodFlow Com- pounds (50 μl, 1%) 30 min 60 min 120 min 180 min Vehicle100^(a) 100^(b) 100^(c) 100^(d) DN-6   56.9 ± 17.5^(e) 61.8 + 20.9* 88.3± 34.1* 128.5 ± 60.0* DN-7 42.0 + 13.7 57.2 + 3.7*  80.5 ± 23.1*  87.3 +29.6* DN-12 62.9 ± 29.6 88.1 + 37.5* 88.9 ± 32.9* 32.2 + 15.0 DN-13105.8 ± 30.1* 83.8 ± 11.3* 153.3 ± 51.5*  112.5 ± 34.6* DC-5  71.4 ±33.3* 71.4 ± 33.3* 80.3 ± 23.7* 124.4 ± 25.4* *Significantly higher thancorresponding controls at 100% ^(a)6.9 ± 3.8 μl/min/mg; ^(b)7.0 ± 2.9μl/min/mg; ^(c)5.6 ± 1.7 μl/min/mg; ^(d)4.9 ± 1.9 μl/min/mg; ^(e)mean ±SD with N = 6 for all except N = 10 for controls.

Since these compounds are able to increase choroidal blood floweffectively, it is expected that they can suppress the triggering effectof choroidal ischemia to induce AMD.

Retinal Function Recovery After Ischemic Insult. When the blood flow tothe retina was cut off for 30 min, the b-wave of the electoretinogramdisappeared, indicating the retinal function was lost. After reperfusionof the retinal blood vessels, however, the b-wave recovered partially toapproximately 30-35% of the original level (Tables 3 and 4). The retinalfunction recovery can be enhanced/facilitated by flavonoids, includingnaringenin, apigenin and puerarin (Table 3). Among them puerarin showedthe best result by showing 90% of retinal function recovery as comparedto control recovery of only 31% (Table 3).

TABLE 3 Effects of Flavonoids on Retinal Function Recovery AfterIschemic Insult Compounds Net (10 mg/kg ip) Control Treated increase (%)Naringenin 32.5 ± 7.5 54.0 ± 17.8* 66.1 Apingenin 32.5 ± 7.5 63.8 ±12.5* 96.3 Puerarin 31.0 ± 9.5 90.0 ± 17.2* 190.3 *Significantly higherthan corresponding controls at P < 0.05 with N = 6 and Mean + SD

N-Nitropyrazoles and C-nytropyrazoles also showed strong retinalfunction recovery after ischemic insult (Table 4). In generalN-nitropyrazoles were more potent than C-nitropyrazoles tofacilitate/enhance retinal function recovery after ischemic insult,except DC-5 which showed stronger effects than any otherN-nitropyrazoles to facilitate the retinal function recovery afterischemic insult (Table 4).

TABLE 4 Effects of N-Nitropyrazoles and C-Nitropyrazoles on RetinalFunction Recovery After Ischemic Insult Compounds Net (10 mg/kg ip)Control Treated increase (%) DN-6 32.9 ± 14.2 64.0 ± 24.9* 94.5 DN-732.9 ± 14.2 71.4 ± 20.2* 117.0 DN-12 32.9 ± 14.2 58.2 ± 16.7* 76.9 DN-1335.0 ± 13.1 58.6 ± 21.4* 67.4 DC-5 33.0 ± 8.6  73.0 ± 16.0* 120.6*Significantly higher than corresponding controls at P < 0.05 with N = 6and Mean ± SD

These results indicate that they are potent agents to enhance retinalfunction recovery possibly though the increase of retinal and choroidalblood flow and are beneficial for the treatment of AMD.

Prevention of CNV Formation

Four weeks after laser treatment of rat's retina to break Bruch'smembrane, choroidal neovascularization was formed and was visible withfluorescein angiography as controls.

A number of agents which can improve choroidal blood flow wereadministered intraperitoneally once a day for 4 weeks. The first type ofagents tested were hypotensive agents including hydralazine, guanabenz,and D-Timolol. Hydralazine at 10 mg/kg inhibited all CNV formationeffectively. When the dose of hydralazine was reduced to 5 mg/kg, onlyfour out of eight CNV formation were clearly inhibited as compared tothe control. Guanabenz was equally effective at 20 mg/kg i.p. All eightCNV formations were inhibited by guanabenz as compared to the control.D-Timolol at 15 mg/kg i.p. was effective in inhibiting some CNVformation but not as potent as hydralazine or guanabenz.

Another group of agents tested was flavonoids, including but not limitedto apigenin, naringenin, quercetin, and flavon. Apigenin at 30 mg/kgi.p. showed a potent inhibition on at least six out of eight CNVformations. Naringenin (30 mg/kg i.p.), quercetin (30 mg/kg i.p.), andflavon (20 mg/kg i.p.) showed marked inhibition of CNV formation. Flavonwas particularly potent and almost completely inhibited the CNVformation.

Another group of agents tested included N-nitropyrazoles (DN's) whichrelease NO to cause vasodilation and choroid blood flow facilitation.All agents, including DN6, DN7 and DN13 at 20 mg/kg i.p. caused markedinhibition of the CNV formation.

Ischemia of choroidal blood flow is closely related to theinduction/triggering of AMD. Therefore, agents that are able to increasechoroidal blood flow will be able to prevent choroidal ischemia and thetriggering of AMD.

It is known that choroidal ischemia can cause RPE cell degeneration,lipofusion accumulation, lyposomal activity changes and extracellularmatrix formation. Further, Bruch's membrane shows abnormality,thickening, and decreases in permeability that leads to the breakdown ofthe Bruch's membrane and the growth of CNV into the subfoveal areas. Asa result visual activity suffered and the AMD developed which leads tothe blindness. Therefore, agents that can increase choroidal blood flow(Tables 1 and 2) should be able to prevent/treat AMD from continuing todevelop, if they are administered at the early stage of AMD.

The effectiveness of these agents to prevent/treat AMD was furthersupported by the fact that they can facilitate/enhance retinal andchoroidal function recovery after ischemic insult (Tables 3 and 4).Therefore, these agents can be used to treat/prevent AMID formation atthe early stage of the disease.

The direct evidence of these agents to prevent/treat AMD comes from theAMD rat model that develop CNV by breakdown of Bruck's membrane by laserbeam. Treatment of these AMD animal model with 1) hypotensive agentssuch as hydralazine, guanabenz, and D-timolol; 2) flavonoids includingapigenin, naringenin, quercetin, and flavon; and 3) N-nitropyrazolederivatives such as DN-6, DN-7, DN-13 at various dose levels showedpotent inhibition of CNV formation which is the main etiology of AMDformation.

In preferred embodiments, agents as IL-1 blockers are antagonists ofIL-1 induced uveitis. Intravitreal injection of IL-1 induced ocularinflammation causes breakdown of the blood-aqueous barrier. As a result,fluorescein may cross the broken blood-aqueous barrier to enter the eyeand to reach peak inflammation 12 hr after IL-1 injection. IL-1 is themost potent cytokine to induce inflammation which can be blockedeffectively by prednisolone at 20 mg/kg t.i.d. (FIG. 13). AmongCK-compounds studied, 9 compounds showed potent blocking effects on IL-1induced uveitis. Although the dose (10 mg/kg t.i.d.) of these compoundswas only half of that of prednisolone (20 mg/kg t.i.d.) used, theyproduced a similar level of IL-1 blockade as compared to prednisolone(FIGS. 12 and 13). CK-120 was particularly potent to block IL-1 induceduveitis as can be seen from FIG. 12.

Some natural products isolated from Chinese herbs were also quite potentto inhibit IL-1 induced uveitis (FIG. 13). Although tetrandrine isslightly less potent than osthol, it is more important because itproduces much less side effects and can be safer when it is usedclinically. Therefore, these agents can be used to stabilize Bruch'smembrane and to prevent CNV and AMD development.

In preferred embodiments, IL-2 blockers delay trabeculectomy failurecaused by Inflammation. Trabeculectomy is used frequently for thetreatment of narrow/closed angle glaucoma. However, it causesinflammation and failure of aqueous humor drainage after a short periodof time after surgery. The synthetic IL-1 blockers such as CK-17,CK-101A and CK-103A prolonged the appearance of trabeculectomy failurethrough inhibition of inflammation caused by IL-1 released from surgery(FIG. 14). They were all more effective than prednisolone to delay thetrabeculectomy failure (FIG. 14). Thus, these agents could be used toprevent breakdown of Bruch's membrane and the development of CNV andAMD.

In preferred embodiments, IL-1 blockers inhibition of systemicinflammation induced by carrageenin. Carrageenin is a potentinflammatory agent to cause pain and swelling on the joints. IL-1blockers such as CK-17 was very effective in inhibiting carrageenininduced inflammation (FIG. 15). CK-17 was about 10 times as potent asaspirin which is a standard agent used widely as anti-inflammatory,analgesic, and anti-arthritis drug (Table 15). These results indicatethat IL-1 blockers can be used locally as well as systematically for theprevention of inflammation and thus beneficial for the inhibition of CNVformation and AMD development.

In preferred embodiments, IL-1 blockers are therapeutically safe. TheLD₅₀ of both CK-17 was extremely high, at least 20 g/kg orally, which isequivalent to 1,400 g/70 kg for man. Since the ED₅₀ of these compoundswas approximately 10 mg/kg, the therapeutic index (LD₅₀/ED₅₀) would behigher than 2,000 (20,000 mg/kg±10 mg/kg). One of the safest agentsavailable.

In preferred embodiment, 1L-1 blockers provide negligible eyeirritation. The ocular irritation of CK compounds was very low andnegligible as can be seen from the Draize test (FIG. 15). This isparticularly important because the agents are to be used for thetreatment of eye disease, AMD.

In preferred embodiment, IL-1 Mocker suppress of CNV formation in rateyes. When the Bruch's membrane was broken by laser beam, massive CNVwas formed with marked fluorescein leakage in the fluoresceinangiography as a control. When the animals were treated by prednisoloneat 3 mg/kg i.p. the CNV formation was markedly inhibited five out ofeight CNV formations.

CK-17 at 30 mg/kg i.p. showed even better results than prednisolone at 3mg/kg i.p. by inhibiting at least six out of eight CNV spots (FIG. 5).The results of CK-112 at 10 mg/kg i.p. were about the same asprednisolone at 3 mg/kg i.p. (FIGS. 4 and 6). When the dose of CK-112was raised to 30 mg/kg i.p. the results were even better (FIG. 7) andwere about the same as that of CK-17 at 30 mg/kg i.p. (FIG. 5). Theeffect of CK-113 at 30 mg/kg i.p. was similar to that of CK-112 at 10mg/kg i.p. (FIGS. 7 and 8).

The effect of CK-115 at 10 mg/kg i.p. was quite impressive (FIG. 9). Itinhibited markedly seven out of eight CNV spots. CK-116 at 10 mg/kg i.p.(FIG. 10) was similar to that of CK-112 at 10 mg/kg i.p. (FIG. 6)whereas CK-117 at 10 mg/kg i.p. was less efficacious (FIG. 11).

However, by raising the doses, the inhibitory action on CNV formationwould be definitely improved, as long as the toxicities of thesecompounds remain at minimal level.

Experiments were done by using laser beam induced CNV through breakingdown of Bruch's membrane. Prednisolone showed inhibition of CNVformation as expected. However, the toxicity of prednisolone and othersteroidal anti-inflammatory agents provides concern for use in theclinics for the treatment of AMD. In this research, 3 mg/kg i.p. ofprednisolone was the minimal dose required to produce inhibition of CNVformation. Yet with this dose level, some animals lost appetite and bodyweight. Higher doses even resulted in the death of animals. On the otherhand, CK compounds were quite non-toxic. For example, CK-17 at 20 g/kggiven orally did not cause any toxic response for at least 7 daysobserved whereas the effective dose to inhibit CNV formation was as lowas 30 mg/kg i.p. Therefore, CK compounds can be used effectively andsafely for the treatment of AMD through CNV formation inhibition.

It is under stood that the compounds can be extracted from naturalsources, purchased commercially, or made by a variety of methods usingprocesses as described in the book by Smith and March “March's AdvanceOrganic Chemistry, 5^(th) edition 2001.” Flavonoids, such as naringenin,apigenin, quercetin, and flavon were commercially available. Hypotensiveagents such as D-Timolol, hydralazine, and guanabenz were purchasedcommercially. Statistical Analysis. All data were analyzed withnon-paired student's t-test. Significance between two means at a certaintime point was considered significant when P≦0.05.

EXAMPLES Example 1 Synthesis of Nitropyrazoles

It is under stood that the compounds can be made by a variety of methodsusing processes as described in the book by Smith and March “March'sAdvance Organic Chemistry, 5^(th) edition 2001”. DC-5, C-nitropyrazoles,were prepared using procedures described in Shevelev et al., Russ ChemBull 44:1861-4 (1993), Kanishchev et al., Bull Acad Sci USSR Div ShemSci 35:2191 (1986), Torf et al., J Gen Chem 32:1740-6 (1962). CompoundsDN-4 through DN-15 were obtained by N-nitration of the correspondingpyrazoles. Compound DN-5 was synthesized according to the methodsprovided in Ugrak et al., Russ. Chem. Bull. 42:1555-1558, 1993. CompoundDN-7 was synthesized according the methods provided in Dalinger et al.,Russ. Chem. Bull. 36:1149-1153, 1997. Compounds DN-8, DN-10, DN-11,DN-12, DN-13, and DN-14 were synthesized according to the methodsprovided in Huttel Chem. Ber. Bd. 88:1586-1590 (1955) and Chem. Ber.88:1577-85 (1955). DN-9 and DN-15 were synthesized according to themethods provided in Janssen el at, J. Org. Chem. 38:1777-1782 (1973).Compounds DN-4 and DN-6 were synthesized according to the methodsprovided in Xuan et al., J. Ocular Pharma. Thera. 17(6):505-515 (2001).

Example 2 Synthesis of Pyridazino[4,5-c]pyridazinones

Synthetic compounds including CK-17, CK101A, CK103A, CK 112, CK 113, CK114, CK 115, CK 116, CK 119, CK 120, and CK 122 were synthesizedaccording to the methods provided in Okawara, et al., Chem. Pharm. Bull.31:507-512. (1983), Yamasaki et al., J. Chem. Soc. Perkin Trans.1:991-996 (1991), Yamasaki et al., J. Heterocyclic Chem. 29:1313-1316(1992), and Bo & Chiou Zhongguo Yao Li Xue Bao. 19(4):304-8 (1998).Other compounds such as flufenamin acid, indomethacin, ibuprofen, andNDGA (Nordihydroguaiaretic acid) and natural products, such asprednisolone, tetrandrine, quercetin, pulegone, friedelin, naringen anddihydrojasmon were purchased commercially. One prepares otherderivatives according to Robins et al., Synthesis of some 7- and5,7-substituted pyrazolo[4,3-d]pyrimidines. J Am Chem Soc 78:2418-22(1956).

Example 3 Measurement of Choroidal Blood Flow in Ocular HypertensiveRabbit Eyes

New Zealand white rabbits, weighing 2.5-3.0 kg, were anesthetized with35 mg/kg ketamine and 5 mg/kg xylazine intramuscularly. Half of theinitial dose was given hourly to maintain anesthesia. An ocularhypertensive model was created by raising the intraocular pressure ofthe left eye to 40 mmHg which reduced the ocular blood flow toapproximately ⅓ of the normal values. The left ventricle was cannulatedthrough the right carotid artery for the injection of microspheres, andthe femoral artery was cannulated for blood sampling. One percent drugsolution (50 μl) or vehicle (50 μl) was instilled topically to the lefteye, and the ocular blood flow of the ocular hypertensive rabbits wasmeasured with colored microspheres at 0, 30, 60, and 120 min thereafter.At each time point, 2 million microspheres in 0.2 mL were injected as areference, and blood samples were taken from the femoral artery forexactly one minute following injection of the microspheres. The bloodsample was collected in a heparinized tube, and the volume was recorded.The rabbits were euthanized with an injection of 100 mg/kg pentobarbitalsodium after the last blood sampling. The left eyes were enucleated anddissected into the retina, choroid, iris, and ciliary body. The tissuesamples were weighed.

The details of sample processing and microsphere counting were providedby E-Z Trac (Los Angeles, Calif.). In brief, Hemolysis Reagent was addedto the microfuge tubes with the blood sample, then vortexed andcentrifuged for 30 min at 6000 rpm. The supernatant was removed, and theTissue/Blood Digest Reagents I and II were added. The tubes were capped,vortexed, and centrifuged for 30 min again. The supernatant was removed,and the Counting Reagent was added, then vortexed, and centrifuged for15 min at the same revolutions as above. The supernatant was removed,and the microspheres were resuspended in a precise volume of theCounting Reagent. The number of microspheres was counted with ahemocytometer.

The Tissue/Blood Digest Reagent I was added to the microfuge tubes withthe tissue samples, sealed, and heated at 95° C. for 15 min. The tubeswere vortexed for 30 sec, reheated and revortexed until all tissuesamples were dissolved. The Tissue/Blood Digest Reagent II was addedwhile the tissue samples were still hot, then the tubes were capped,vortexed, and centrifuged for 30 min. The protocol, thereafter, was thesame as that used to process the blood sample, and the microspheres werecounted.

The blood flow of each tissue at a certain time point was calculatedfrom the following equation:

Qm=(Cm×Qr)/Cr

Where Qm is the blood flow of a tissue in terms of μL/min/mg, Cm is themicrosphere count per mg of tissue, Qr is the flow rate of blood samplein terms of μL/min, and Cr is the total microsphere count in thereferenced blood sample.

Example 4 Measurement of Retinal Function Recovery After Ischemic Insultin Rat Eyes

Electroretinograms (ERGs) were determined to provide assessment of theretinal function prior to and following ischemic insult. ERGs wererecorded by means of Ag/AgCl electrodes placed in contact with thecornea. One stainless steel needle was inserted subcutaneously betweenthe two eyes as a reference electrode, and another needle was insertedsubcutaneously to the neck as a ground electrode. A photostimulator(Grass PS22 Flash) was used to produce flashes of light five inches fromthe eye, and the ERG potentials were recorded with a polygraph system.The ERG machine was purchased from LKC Technologies, Inc. (Gaithersburg,Md.). A single (10 msec duration), white light stimuli was used toelicit ERG a- and b-waves. Peak b-wave amplitudes were measured from thetrough of a-wave to the peak of b-wave.

Dark-adapted, female Long-Evans rats (200-250 g) were anesthetized with35 mg/kg ketamine plus 5 mg/kg xylazine intramuscularly. Half of theinitial dose was given thereafter at one-hour intervals to maintainadequate anesthesia. The pupils were dilated with 1% tropicamide plus10% phenylephrine (50 μl) for ERG experiments. Retinal ischemia wasproduced by occlusion of the central retina and posterior ciliaryarteries by means of a ligature placed around the optic nerve and theposterior ciliary artery. The ligature was then tightly drawn for 30 minto occlude the retinal vessels. Retinal ischemia was confirmed by theextinction of the ERG waves. After 30 min of retinal ischemia, ligaturewas released and the retinal arteries allowed to reperfuse. ERGs werethen measured at 0, 30, 60, 90, 120, 180, and 240 min thereafter. Drugsand vehicles were administered intraperitoneally. These drugs wereadministered immediately prior to occlusion of the central retinalarteries.

Example 5 Measurement of CNV Formation in Rat Eyes after Induction ofChoroidal Neovascularization

Brown-Norway rats, weighing 200˜250 g, were anesthetized with ketamine35 mg/kg and xylazine 5 mg/kg. Pupils were dilated with a topicalapplication of 1% tropicamide (Bausch & Lomb; Tampa, Fla.) and 2.5%Neo-Synephrine (Sanofi-Synthelabo Inc.; NY). CNV was experimentallyinduced with a double frequency Nd:YAG laser (Laserex LP3532; EllexMedical PTY. LTD., Australia) to disrupt Bruch's membrane. The laserwavelength was 532 nm, and spot size was 100 μm. Power delivered rangedfrom 130 to 150 mW, applied for 0.1 sec. Typically, eight lesions wereinduced in both eyes of each animal. On occasion, the inducing laserburst created an extensive subretinal and/or vitreous hemorrhage, andthese spots were excluded from any further treatment or analysis. Otherlesions in the same eye were included in the study if the subretinalhemorrhage did not extend to within approximately 1 mm of the lesion.The presence of CNV was confirmed at 2 weeks after laser induction byfluorescein angiography. Each laser lesion was evaluated as follows:Leaking (++): increase in size of hyperfluorescence over time;Non-leaking (+): only staining without increase in size ofhyperfluorescence; Non-evaluation (−): fluorescein angiography could notevaluate the lesion due to masking by the overlying hemorrhage.

Fluorescein Angiography. Fluorescein angiography was performed inanesthetized animals with dilated pupils using a Digital Fundus Camera(TRC-50 EX: Topcon, Japan) and standard fluorescein filter 0.3 ml of 10%fluorescein (Sigma-Aldrich Inc.; St. Louis, Mo.) was injectedintravenously via hypoglossal vein. After injection, the cameraalternated between both eyes taking images for 10 minutes. Fluoresceinangiography was performed in all rats at 2 and 4 weeks followingsimultaneous laser induction of CNV and drug-vehicle injection.

In vivo Evaluation: All rats were examined weekly after laser inductionof CNV and drug/vehicle injection up to the time of sacrifice. Theexamination was performed by slit lamp (SL-3E; Topcon, Japan) toevaluate the anterior segment and by indirect ophthalmoscope (Omega 200;Heine, Germany) using a 20 D lens to evaluate the retina and vitreous.

Example 7 Interleukin-1 Induced Uveitis

Female Sprague-Dawley rats, weighing 250-300 g, were anesthetized with35 mg/kg ketamine and 5 mg/kg xylazine intramuscularly. Ten μl of 1.0 ngof IL-1α were injected intravitreously with 30-gauge needle, and therats were allowed to recover from the anesthesia. CK-compounds, at dosesof 3 mg/kg or 10 mg/kg, were injected intraperitoneally (i.p) at time 0,4, and 10 hours after the IL-1α injection. The inflammation was measured12 hours after IL-1α injection. The rats were anesthetized again asdescribed previously, and FD-70 solution (1.4 ml/kg) was injectedintravenously through the hypoglossal vein. Scanning of the eyes wasconducted with a fluorophotometer (Fluorotron Master, Coherent Corp.,Palo Alto, Calif.). Measurements were carried out at 0, 30, 60, 90, 120,180, 240, 300, and 360 min after FD-70 injection. The peak level ofFD-70 in the eye was reached at 300 min after the FD-70 injection.

Example 8 Trabeculectomy Induced Inflammation

Adult Dutch Belted rabbits were purchased commercially and were specificpathogen free for Pasteurellosis. Rabbits were housed in individualcages and kept on twelve-hour, light-dark cycle. The room temperaturewas maintained at 25° C. and the relative humidity was 50%. Rabbits weregiven access to rabbit chow (Harland, Houston, Tex.) and water adlibitum. Rabbits were fasted for twelve hours before induction ofanesthesia.

The animals were predosed with an antimuscarinic and sedative. Therabbit was induced with ketamine/xylazine, intuvated and then maintainedanesthetized with isoflurane. Buprenorphine and pancuronium were alsoused for a balanced anesthetic regimen. Ten mg/kg buprenorphine wasadministered every 12 hours for three days for analgesia postoperatively.

A glaucoma filtration operation with partial thickness scleral flap wascompleted using an operating microscope and a strict aseptic technique.A limbus based conjunctival flap was created. Then, approximately onemillimeter temporal to the superior rectus, a partial thickness miniflapwas formed in the sclera. The flap dimensions were three millimeters atthe limbus, two millimeters on the sides and at the base. At thesurgical limbus a 2.5 millimeter sclerostomy was performed using a razorblade chip. A Kelly Descement punch (Storz, St. Louis, Mo.) was thenused to complete the sclerostomy. A peripheral iridectomy was thenperformed. The iris was reposited with a balanced salt solution flushand gentle corneal massage. The scleral flap was then sutured closedwith a single 10-0 micorsurgical nylon suture and fixed with anoverlying square knot. The conjunctiva and the tenons incision wereclosed with a running 10-0 suture. Immediately upon water-tight closureof the conjunctiva, a filtration bleb could be noted. The same procedurewas done on the contralateral eye.

In vivo Evaluation. All rats were examined weekly after laser inductionof CNV and drug/vehicle injection up to the time of sacrifice. Theexamination was performed by slit lamp (SL-3E; Topcon, Japan) toevaluate the anterior segment and by indirect ophthalmoscope (Omega 200;Heine, Germany) using a 20 D lens to evaluate the retina and vitreous.

Prior to surgery, the rabbits were randomly assigned to one of sevengroups 1) vehicle control subtenons injection (STI), 2) prednisolone eyedrops, 3) no drug 4) methylprednisolone STI, 5) CK-17 STI, 6) CK-101ASTI, and 7) CK-102 STI. The animals in the eye drops group received twodrops in each eye three times per day. The eye drops 50 μl treatmentbegan the day before surgery and continued until failure of the fistula.The control for the eye drops group was “no drug” because theformulation of the eye drops is patented and therefore, a vehiclecontrol could not be prepared. If the animal was in subtenons treatmentgroup, then 10 mg of the drug was given by subtenons injection while theanimal was still under general anesthesia. If the animal was in thevehicle control group, a volume of the vehicle was given equal to thatgiven in the treatment groups. Finally, 20 mg gentamicin was givensubconjunctivally for prophylaxis of microbial infection.

The eyes were preanesthetized with one to two drops of tetracaineinstilled into both eyes and the intraocular pressure was measured byapplanation pneumotonography (Alcon, Ft. Worth, Tex.). Preoperativemeasurements were recorded the day before surgery. Postoperativemeasurements were recorded every other day. Failure of the filtrationfistula was defined as the point where IOP returned to preoperativevalues or the same value was recorded for three consecutive readings.The presence or absence of the filtration bleb was also recorded everyother day. On the days when IOP was not measured, biomicroscopeexaminations were conducted and the inflammation was scored using thesystem previously defined by Miyano & Chiou, Ophthalmic. Res. 16:256-263(1984). For biomicroscope examination, the rabbit was anesthetized with1 mg/kg acepromazine IM. Inflammation scores were calculated andrecorded. At the conclusion of the experiment the animals wereeuthanized with an overdose (100 mg/kg) of pentobarbital sodium.

Example 9 Carrageenin-Induced Inflammation

Forty female Sprague Dowley rats weighing 164±16 g were used for theexperiments. The test and control groups were assigned randomly.Carrageenin (0.1 mL of 1% solution) was injected into the plantarsurface of the rat hind paw to induce inflammation. Ten minutes beforecarrageenin injection, 8 mL of water administered ig. CK-17, and aspirinwere grounded with Tween 80 and then suspended into distilled water.CK-17, or aspirin were injected ip 3 times at time 0, 8, and 16 h beforethe injection of carrageenin. The changes in volume of the inflamededematous foot were measured by the volume of water displaced andrecorded at 0, 0.5, 1, 2, 4, and 6 h after the carrageenin injection.

LD₅₀ determination: For each dose, 20 mice of either sex, weighing 18-20g were used to determined the LD₅₀ according to the method of Litchfield& Wilcoxon, J. Pharmacol. Exp. Ther. 96:99-113 (1949). Animals werehoused in an animal room at 25° C. and 70% relative humidity. The lightcycle was set for 12 h light and 12 h darkness. CK-17 was grounded withTween 80 and then suspended in 1% CMC (carboxymethyl cellulose). Thesuspension was administered ig at 20 g/kg and the animals were observedfor 7 d.

Example 10 Eye Irritation Test

Draize test, Draize et al., J. Pharmacol. Exp. Ther. 82:377-90 (1944),was followed for the determination of eye irritation. Before the Draizetest, it was already known that CK-17 did not produce skin irritation inguinea pigs. So, Draize test was used to show the safety rather than thetoxicity of drugs in the rabbit eyes in this experiment. Six New Zealandalbino rabbits of either sex, weighing 2-2.5 kg were used in each group.Animals were housed individually in cages at 25° C. and 70% relativehumidity. The light cycle was maintained at 12 h light and 12 h dark for7 d after the instillation of CK-17 or vehicle as controls.

CK-17 (0.1%) was suspended in 6% dimethylsulfoxide (DMSO), 6% PEG 400,10% Tween 80, and 78% saline. Fifty microliters of the test compound wasinstilled into the cul de sac of the right eye as treated and thevehicle to the left eye as control. The eye lids were gently heldtogether for 10 s to prevent the loss of the materials. Animals showingeye irritation, ocular defects or pre-existing corneal injuries were notused for the experiment. The eyes were examined with ophthalmoscopy at0, 1, 3, 5, 7, and 24 h, and 2 d, 3 d, 5 d, and 7 d after the eye dropinstillation. The scores of the Irritation Table were recorded andcalculated.

All publications and patent applications mentioned in the specificationare indicative of the level of skill of those skilled in the art towhich this invention pertains. All publications and patent applicationsare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

All of the compositions and/or methods disclosed and claimed herein canbe made and executed without undue experimentation in light of thepresent disclosure. While the compositions and methods of this inventionhave been described in terms of preferred embodiments, it will beapparent to those of skill in the art that variations may be applied tothe compositions and/or methods and in the steps or in the sequence ofsteps of the method described herein without departing from the concept,spirit and scope of the invention. More specifically, it will beapparent that certain agents which are both chemically andphysiologically related may be substituted for the agents describedherein while the same or similar results would be achieved. All suchsimilar substitutes and modifications apparent to those skilled in theart are deemed to be within the spirit, scope and concept of theinvention as defined by the appended claims.

1. A method of treating macular degeneration, comprising: topicallyadministering a composition comprising hydralazine or a salt thereof toat least one eye of a subject diagnosed with age-related dry maculardegeneration.
 2. The method of claim 1, wherein said subject is a human.3. The method of claim 1, wherein said salt is a hydrochloride salt, ahydrochlorothiazide salt or an isosorbide dinitrate salt.
 4. The methodof claim 1, wherein said composition is a liquid solution.
 5. The methodof claim 1, wherein said administering comprises topically administeringthe composition to both eyes of the subject.
 6. A method of treatment,comprising: topically administering a composition comprising hydralazineor a salt thereof to at least one eye of a subject diagnosed withage-related dry macular degeneration and at risk for choroidalneovascularization.